It has been highlighted that shame may be an important dynamic risk factor for prevention of violence and recidivism in forensic populations. However, past research investigating the relationship between shame and violence, or recidivism has been inconsistent. Different conceptualisations and measurements of shame used in the literature may explain these inconsistencies. Therefore, a systematic review was conducted to explore how shame was conceptualised in forensic populations and these measures were then evaluated. Findings revealed that most studies did not clearly define shame, and when they did, the same theoretical underpinnings were used in different ways. By assessing the validity and reliability of shame measures, it was revealed that different measures focused on different aspects of shame. This could explain the current confusion in the conceptualisation and measurement of shame in forensic populations, and shed light on inconsistent findings between shame and other constructs. Shame in violent female offenders is an unexplored phenomenon and therefore may involve various complex and unexpected factors. A social constructivist grounded theory approach was applied to the narratives of eight violent female offenders, focusing on thoughts, feelings and life experiences in relation to shame and violence. A model was constructed suggesting that childhood victimisation, in the absence of available, compassionate, secure relationships, may lead to difficulties with emotion regulation. The experience of negative emotions, including shame, may lead to self-harm, substance misuse and violence. It was however demonstrated that this vicious cycle could be broken through the development of secure, positive and compassionate relationships. These findings suggest that shame and attachment may be important factors for treatment and service planning, to meet the unique needs of female offenders.

The high mobility group protein HMGA2 is an architectural transcription factor, which is expressed during embryogenesis. Aberrant expression causes benign and malignant tumor formation. The protein possesses three "AT hook" domains and an acidic Cterminal. HMGA2 is natively unstructured, however it forms a homodimer. In this study site-directed mutagenesis was used to create single methionine mutants, HMGA2Q37M, HMGA2I71M and HMGA2Q85M. These mutants were cross-linked using EDC and then cleaved using CNBr to determine which domains are involved in homodimer formation. Our results indicate that the second "AT hook" domain may interact with the C-terminal. We then labeled a peptide containing the C-terminal (CTP) with tetramethylrhodamine-5- maleimide (TRM). We found that the CTP-TMR binds to HMGA2Α95-108, which lacks the C-terminal. These results suggest that the C-terminal is required for homodimer formation. The techniques used within this study can be applied to forensics and with further research HMGA2 may have a forensic application.

Due to the threat of bioterrorist acts, there is a need to develop techniques that rapidly detect possible bioagents. Amplicon length heterogeneity-polymerase chain reaction (ALH-PCR) presumptively identifies eubacteria in samples by detecting differences between the lengths of the hypervariable regions of the 16S rRNA gene. To study the efficiency, reproducibility, and reliability of the technique, sputum from cystic fibrosis (CF) patients has been chosen as the model system. There is an abundance of microorganisms in the sputum of the CF lung. Using ALH-PCR, the complex microbial diversity and vast community composition in the lungs of the CF patients were studied. Twenty-four out of twenty-six CF samples were presumptively identified to contain Pseudomonas aeurginosa, a known CF pathogen. Sputum profiles were also compared over time and ALH was able to demonstrate that the CF lung flora is a dynamic community and may be affected by antibiotics.

The purpose of this research was to develop a strategy for the DNA typing of Cannabis sativa. DNA typing was accomplished by using the Amplified Fragment Length Polymorphism (AFLP) method. Cannabis (marijuana) was selected because it is an illegal substance grown across the United States and in other countries and the sourcing of this plant is of interest to law enforcement agencies. While some peaks may be shared, the overall peak profile is expected to contain some peak differences between individuals. The AFLP procedure was performed on 25 different cannabis samples using four different primer combinations. In all of the AFLP profiles, there were significant peak differences that allowed for all the samples to be distinguished. The samples that were genetically related displayed the same peak profiles in the electropherograms and the samples that were not related possessed unique profiles. The development and validation of the AFLP method would lead to the creation of a database that could then be used to link cloned samples and to track distribution networks and, ultimately, individual plants could be linked together.

The purpose of this research is to introduce a method for the forensic elemental analysis of cotton fibers for the purpose of increasing the discrimination between otherwise similar cotton evidence using microwave digestion Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) and Laser Ablation- Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS). A quadrupole ICP-MS and UV laser ablation (266nm) instruments were used for the analysis. A cotton standard reference material (IAEA V-9) was used to validate the developed methods producing good accuracy with typically 10 % bias and good precision (typically 5% RSD) for the element list: 25Mg, 27Al, 55Mn, 57Fe, 88Sr and 137Ba. It was found that the LA-ICP-MS method resulted in improved precision over the solution ICP-MS method. Twenty four (24) raw cotton samples and five white cotton T-Shirts were analyzed with the developed methods. It was also found that all the raw cotton samples from different sources were distinguishable from each other, as were all the cotton T-shirts resulting in zero type I errors and zero type II errors for the pairwise comparisons.

I. Investigations were carried out to establish (a) the regulation of magnesium homeostasis in the exocrine pancreas under agonist stimulation (b) that secretagogueevoked cellular processes in the exocrine pancreas are sensitive to physiological changes in Mg2+ and (c) the intracellular roles of Mg2+ in the secretagogue-evoked exocytotic processes in the exocrine pancreas. H. CCK8 and ACh can induce a dose-dependent transmembrane efflux of Mg2+ which results in a subsequent lowering of [M g2+] . The routes for movement of Mg2+ do not appear to be associated with those sites of Ca2+ mobilisation from either the internal Ca2+ stores or across cell membranes but appears to be associated with those of Na+ and is ATP-dependent. Stimulation with secretin results in an elevation of [Mg2+] compared to resting values. The secretin-evoked elevation of [Mg2+] may be coupled to G protein activation of adenylate cyclase. Digital imaging technology further demonstrates that mobilisation of Mg2+ follows a different route from that of Ca2 . The CCK8-evoked Ca2 signal is initiated in an area towards the luminal pole of the pancreatic acini and rapidly spreads throughout the entire cell. Stimulation of single pancreatic acini with CCK8 results in an initial transient increase in [Mg 2 ] throughout the entire cell which is followed by a sustained decrease in [Mg2+] which is more prominent around the basolateral membrane. III. Pertubations in extracellular and intracellular Mg2+ had marked effects on the CCK5- and secretin-evoked pancreatic juice flow and protein output in (a) the isolated perfused intact pancreas, and (b) amylase release in pancreatic segments and permeabilised pancreatic acini. A nominally Mg2+ deficient saline augments the CCK8-induced responses, whereas elevated Mg2+ attenuated secretion compared to normal (1.0 mM) Mg2 conditions. Parallel effects were found on Ca 2 mobilisation in Fura 2 loaded acini suspensions and on oscillations of Ca2+ in single acini stimulated with CCK8. These effects of Mg 2 appear to be mediated by direct ffil modification of Ca2+ release, propagation and re-uptake into cytoplasmic stores. The effects of Mg2+ on secretin-evoked secretion followed a bell shape characteristics curve for Mg2+dependent enzyme activity, i.e. elevating and reducing the intracellular free magnesium levels attenuate the secretin-induced response. IV. Infusion of CCK8 and secretin evoked time course increases in both pancreatic juice flow and total protein output in anaesthetised rats and in the isolated perfused intact pancreas. Simultaneous application of CCK5 and secretin caused some degree of attenuation in secretory responses. This reduction in secretion may not involve protein kinase C or cyclic AMP but may occur at some point prior to the activation of protein kinase A. Secretin may influence mobilisation of Ca2+ which is required to initiate secretion, possibly by stimulating influx of Mg 2 . V. Electrical stimulation of vagus nerves led to elevated pancreatic juice flow and protein output in anaesthetised rats. Secretin attenuated these secretory parameters. Activation of protein kinase C with TPA, had no significant affect on these nervemediated responses. Electrical field stimulation (EPS, 50 V, 20 Hz, 1 msec) andlor ACh stimulation of pancreatic segments results in marked amylase release; 45Ca2+ influx and elevations in [Ca 2+]i. Secretin had little effect on these parameters but attenuated the EPS and ACh-induced responses. The secretin-evoked inhibition of EFS and ACh -induced secretion was abolished when extracellular Mg2+ was absent. Secretin may control nerve-mediated secretory responses by stimulating an influx of Mg2+ to interact with Ca2+. VI. The present investigations have demonstrated that transport mechanisms exist in the exocrine pancreas to regulate cytosolic free Mg2+ levels and that exocytotic processes exist in the exocrine rat pancreas which are sensitive to Mg2+ within a physiological range. Mg2 may act as an intracellular modulator, primarily by controlling mobilisation of Ca2+.

The persistence and detection of gene transcripts in post mortem tissues has significance for forensic science and medicine, since there is the potential to provide information about the cause of, or circumstances surrounding, death and the post mortem interval. Previous studies in this area have focused on enzyme activities and the presence of gene transcripts in neural tissues (Palego et al., 1993; Johnson ci aL, 1986). In the study presented in this thesis, the biological material selected was dermal tissue from pigs, since the skin offers a number of advantages for study in a forensic context, such as ease of harvesting and a slower rate of degradation than most other soft body tissues. Initial chapters describe the development of methodologies for the extraction of RNA from dermal tissue and the detection and quantification of gene transcripts. The genes selected as indicators of gene expression were members of the heat shock gene family, since these genes are highly conserved and easily induced in cells. Subsequent chapters present evidence, not only for the persistence and detection of gene transcripts in post mortem tissue, but also for the inducible de novo expression of heat shock genes in post mortem (PM) tissue. A mild heat shock, applied in the post mortem period to porcine tissue, elicits the expression of certain genes of the heat shock gene family. These results demonstrate that heat shock genes are highly inducible in the early post mortem period, and that these cells retain the ability to respond to an environmental stimulus by transcribing specific genes.

Many eukaryotic organisms produce membrane interactive, a-helical antimicrobial peptides (a-AMPs) and a database of such peptides, together with selected physiochemical parameters was established. This database was divided into four groups according to the a-AMPs target organism(s) (active against Gram-positive bacteria {G+}; active against Gram-negative bacteria {G-}; active against Grampositive and Gram-negative bacteria {G+, G-}; or active against Gram-positive bacteria, Gram-negative bacteria and fungi {G+, G-, F}). Analysis of the database showed that there was no statistically significant correlation between specificity and p1 (range 4.2 to 12.7) or net charge (range -5 to +16). The peptides exhibited variable hydrophobicity, < H > (range -0.8 to +0.7) whilst amphiphilicity (measured by the hydrophobic moment, < jAH >) ranged from 0.2 to 1.1. A statistically significant negative correlation between < pH > and < H > was noted for each group of a-AMPs and this may relate to the amphiphilic balance required for antimicrobial activity, a-AMPs showed some differences in amino acid composition compared to the McCaldon and Argos dataset of unrelated oligopeptides, suggesting functional relevance of some amino acid residues. The groups {G+, G-} and {G+, G-, F}, for example are characterised by being rich in weakly hydrophobic or hydrophilic amino acids. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) can provide a measure of AMP potency. A statistical analysis of MIC's for peptides from {G+, G-} group, showed no significant differences in the potency of these peptides when directed against either Gram-positive or Gram-negative bacteria. In contrast, a statistical analysis of MIC's for peptides from the {G+, 0-, F} group showed that peptides from this group were effective at lower concentrations against bacterial targets as compared to fungal targets. Increases in hydrophobic arc size were generally accompanied by increases in peptide antimicrobial potency and in addition, a negative correlation between MIC and net charge was observed. Regression analyses indicated that an appropriate amphiphilicity/hydrophobicity balance was required for the antimicrobial action of a-AMPs and this may indicate a general structure/function relationship underlying both the efficacy and specificity of these peptides. Oblique orientated a-helices are highly specialised protein structural elements that penetrate membranes at a shallow angle and are used to promote membrane destabilisation by a number of protein classes. Here, the use of extended < pH > methodology showed that over 50% of the a-AMPs are candidate oblique ahelices providing some insight into possible modes of action. Peptides VP I and BYDV-MP were identified here as candidate AMPs based on amino acid composition and the potential to form oblique orientation. The biological activity of VPI and BYDV-MP was confirmed in vivo when an MLC of 3 mM was demonstrated on both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli. Monolayer studies using lipid extract from these target organisms and parallel studies using mimetic monolayers confirmed a high level of peptide membrane interaction. The single lipid monolayer results suggested that VPI and BYDV-MP have a lower affinity for zwitterionic lipid (DMPE surface pressure increase of 4 mNm') but a high affinity for anionic lipid (DMPS surface pressure increase of 7 to 9 mN m') and may have a requirement for this specific lipid or anionic lipids in general to achieve higher levels of membrane penetration. To test the ability of VP! to penetrate membranes protonated and deuterated homologues were analysed by neutron diffraction, in the presence of POPC: POPS (10:1 molar ratio). The data analysed from these studies showed the protonated homologue to penetrate the membrane core but the deuterated homologue showed no significant levels of membrane interaction. Monolayer studies confirmed that the protonated homologue interacted strongly with anionic and zwitterionic membranes (surface pressure increase 4 mNm'), however, the deuterated homologue did not have the ability to interact with POPC:POPS monolayers. FTIR conformational analysis showed the protonated homologue to adopt high levels of a-helical stmcture (65 %) and in contrast the deuterated homologue exhibited low levels of a-helical structure (C 20 %). These results support the original predictions and also appear to show that deuteration has directly or indirectly, affected the ability of the VP! peptide to interact with membranes, possibly by inhibiting a-helix formation by the peptide or decreasing structural stability.

Forged documents are comnonly encountered in the investigation of crime. Forensic document examiners need to be able to employ techniques that are non-destructive, reliable and admissible in court. In this work, techniques currently in use for ink analysis, including FLE, TLC, UV - Vis, IR-ATR and Raman spectroscopy, have been reviewed and, where possible optimised in order to evaluate the level of discrimination possible between different ballpoint and gel pen inks. To date, information comparing such a wide range of techniques to relatively large sample sets has not been published. Particularly, lacking is analysis of ballpoint pen inks from the UK, ballpoint pen ink colours other than black and blue and any analysis of gel pen inks. The results vary widely depending on the combination of ink colour and type, paper and the technique being used. FLE achieved the best discrimination for blue gel pen inks giving a DP of 0.92. TLC is most effective for black and red ballpoint pen inks, giving a DP of 0.89 for both. The highest DP for UV - Vis, 0.86, is achieved for black gel pen inks and for IR-ATR it is red gel pen inks that are most effectively discriminated between, with DPs of 0.88 and 0.87. The Raman spectroscopy analysis used a greater number of combinations of papers, and experimental parameters, i.e. different excitation wavelengths and 'standard' Raman spectroscopy or SERRS with silver or gold colloid, therefore, it is not possible to select a single type and colour of ink to which the technique is best suited. The pairs of inks discriminated by the five techniques are not mutually exclusive. In real casework it is unlikely that all the instrumentation would be available and there will be financial constraints on how many techniques can be carried out. Therefore, using the data obtained here, protocols have been suggested in order to predict the technique or combination of techniques that is most likely to achieve discrimination of two different inks for each pen ink type and colour. Raman spectroscopy or SERRS are required for 23 out of the 28 recommended protocols suggesting that they would be valuable tools for document examination laboratories. However, for Raman spectroscopy, although it is a non-destructive method of analysis, when the SER.RS enhancement technique is used, which is effectively non-destructive, some problems with reproducibility were found. Causes of the irreproducibility have been put forward, such as inhomogeneity of the ink and the paper substrate and ink and paper interactions, but no single cause has been identified. Despite this, if care is taken with analysis and interpretation, SERRS results can be useful.

Based on ethnographic fieldwork and drawing on anthropological, clinical, and social science literature, Without Power, Without Glory examines the work of a palliative care team of physicians, social workers, and nurses who cared for children and families who faced catastrophe and could not be cured in an institution whose professional identity, metrics for success, and global advertising campaigns are centered on cure. This dissertation details the social construction of pediatric palliative care, which is often wrongly seen as both synonymous with hospice and clouded by the team’s close relationships with patients and families. Since the palliative care team often follows patients for many years, it also captures the way that children age out of innocence and into suspicion, especially with regards to pain medication for chronic illnesses. As a consult service that operates at a financial loss for the hospital, this dissertation reports on the palliative care team’s struggle to advocate for patients and families in the face of bureaucratic indifference. Though the hospital aggressively recruits children with complex illnesses for financial gain, it documents how comfort is pitted against technical care, which means the palliative care team and especially chaplaincy services are often excluded or not even hired. Finally, Without Power, Without Glory explores truth and lie in the disclosure of prognosis to families and shows how the responsibility to foresee is often diffused from the medical team onto children who are said to speak through tests. / Anthropology